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Proteomic analysis of RUNX2 overexpressed DFCs in miniature pig

ABSTRACT: We analyzed the features of proteome profiling of the supernatant from dental follicle cells from miniature pigs. The molecular mechanism revealed in this study is of great significance for future studies on the timing of organ renewal or regeneration.

ORGANISM(S): Sus Scrofa Domesticus

SUBMITTER: Songlin Wang

PROVIDER: PXD042486 | iProX | Wed May 24 00:00:00 BST 2023

REPOSITORIES: iProX

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Expression data from early diphyodont morphogenesis in minature pig

Project description:The miniature pig is diphyodont, making it a valuable alternative model for understanding human tooth development and replacement. However, little is known about gene expression and function during swine odontogenesis. The goal of this study is to undertake the survey of differential gene expression profiling with Affymetrix Porcine GeneChip and functional network analysis during morphogenesis of diphyodont dentition in miniature pigs. The identification of genes related to diphyodont development should lead to a better understanding of morphogenetic patterns and the mechanisms of diphyodont replacement in large animal models and humans. The staged miniature pig embryos and fetuses were obtained by cesarean section at E40, E50, and E60. The last deciduous molar germs, including the dental lamina in mandibles from the same litter were isolated and pooled. The gestational age of embryos used in our study just cover 3 characteristic stages: Dm3 in cap stage without secondary dental lamina (E40); Dm3 in bell stage with secondary dental lamina initiation (E50); Dm3 in secretory stage without evidence of morphological changes in the secondary dental lamina.

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Gene expression profiles of DFCs and SHED 48 hours after in vitro transfection with a TP53 plasmid, a SP1 plasmid, or an empty vector.

Project description:Dental follicle is a loose connective tissue that surrounds the developing tooth. Dental follicle cells (DFCs) have a promising potential for tissue engineering applications including periodontal and bone regeneration. However, little is known about the molecular mechanisms underlying osteogenic differentiation. In a previous study we detected that more than 35 % of genes that are regulated during osteogenic differentiation of DFCs have promoter binding sites for the transcription factors TP53 and SP1. However, the role of these transcription factors in dental stem cells is still unknown. We hypothesize that both factors influence the processes of cell proliferation and differentiation in dental stem cells. Therefore, we transiently transfected DFCs and dental pulp stem cells (SHED; Stem cells from human exfoliated decidiuous teeth) with expression vectors for these transcription factors. After overexpression of SP1 and TP53, SP1 influenced cell proliferation and TP53 osteogenic differentiation in both dental cell types. The effects on cell proliferation and differentiation were less pronounced after siRNA mediated silencing of TP53 and SP1. This indicates that the effects we observed after TP53 and SP1 overexpression are indirect and subject of complex regulation. Interestingly, upregulated biological processes in DFCs after TP53-overexpression resemble the downregulated biological processes in SHED after SP1-overexpression. Here, regulated processes are involved in cell motility, wound healing and programmed cell death. In conclusion, our study demonstrates that SP1 and TP53 influence cell proliferation and differentiation and similar biological processes in both SHED and DFCs. Total RNAs were isolated from dental follicle cells after 48 hours of transfection with a TP53 expressions plasmid, a SP1 expressions plasmid and for control with an empty vector.

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Gene expression in feather dermal papilla (DP)

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Comparative gene expression analysis of dental follicle and periodontal ligament in humans

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Comparative bulk RNAseq analysis of Control and PTH/PTHrP recepter-deficient PTHrP+ dental follicle cells

Project description:Lack of PTH/PTHrP receptor in PTHrP+ dental follicle cells causes a cell fate shift and premature differentiation. Here, we set out to reveal the transcriptome change induced by deficiency in PTH/PTHrP receptor in PTHrP+ dental follicle cells

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The cellular basis for epithelial-mesenchymal interaction during mouse tooth development

Project description:In this study, using mouse molar as the model, we developed a dual fluorescence reporter mouse to precisely track and analyze dental epithelium and mesenchyme at single-cell resolution from early embryonic to postnatal stages. Moreover, we constructed the virtual molar explorer (VMEx) to spatially map 15,967 molar-expressed genes and identified that Msx1+ Sdc1+ marked the developing dental papilla while surrounded by Msx1+ Sdc1- molar niche. Through tooth germ reconstitution and organoid culture in vitro and kidney capsule transplantation in vivo, we provided evidence that the Msx1+ Sdc1- dental follicle cells might function as the tooth organizers that promoted epithelium survival and tooth germ organization. Furthermore, the appearance of Msx1+ Sdc1+ dental papilla cells relied on the interaction between dental epithelium and Msx1+ Sdc1- dental follicle cells. Together, our results revealed the cellular dynamics of tooth development in mice and identified that the dental follicle might be the key driver of epithelial-mesenchymal interaction and tooth morphogenesis.

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